The development of organized vascular networks necessitates a tightly regulated interplay between variable cells, growth factors and soluble mediators. The applicant's long-term goal is to develop therapeutic angiogenic strategies based on the rational design of cytokine releasing constructs that promote vascular patterning and vessel stability. The objective of this proposal is to i) develop electrospun, three-dimensional constructs with patterned architecture, ii) demonstrate that the spatial and temporal delivery of two model angiogenic growth factors promotes the formation of an organized capillary network and iii) develop a computational model that can predict the biological effect of a growth factor releasing construct as a function of specified fabrication parameters. We hypothesize that guided therapeutic angiogenesis (i.e. patterned vascular networks) can be obtained by controlling the spatial and temporal presentation of soluble mediators at the site of ischemia. In AIM I, we will synthesize bFGF and G-CSF releasing electrospun constructs and determine their programmed delivery as a function of fabrication parameters. In AIM II, we will demonstrate the effect of spatial and temporal control of cytokine delivery in promoting directed angiogenesis in a three-dimensional in vitro angiogenesis model and we will develop a computational model/software that can predict the biological effect of different scaffold configurations. We will validate our model by assessing the angiogenic potential of our growth factor releasing constructs in a murine critical limb ischemic model. PUBLIC HEALTH RELEVANCE: We are proposing to a) fabricate a growth factor releasing construct that regulates the spatio- temporal delivery of angiogenic cytokines and promotes the formation of organized capillary networks and b) develop a machine learning computational model that can predict the angiogenic potential of our construct as a function of fabrication parameters.